Unknown

Dataset Information

0

Ligand discrimination between active and inactive activation loop conformations of Aurora-A kinase is unmodified by phosphorylation.


ABSTRACT: Structure-based drug design is commonly used to guide the development of potent and specific enzyme inhibitors. Many enzymes - such as protein kinases - adopt multiple conformations, and conformational interconversion is expected to impact on the design of small molecule inhibitors. We measured the dynamic equilibrium between DFG-in-like active and DFG-out-like inactive conformations of the activation loop of unphosphorylated Aurora-A alone, in the presence of the activator TPX2, and in the presence of kinase inhibitors. The unphosphorylated kinase had a shorter residence time of the activation loop in the active conformation and a shift in the position of equilibrium towards the inactive conformation compared with phosphorylated kinase for all conditions measured. Ligand binding was associated with a change in the position of conformational equilibrium which was specific to each ligand and independent of the kinase phosphorylation state. As a consequence of this, the ability of a ligand to discriminate between active and inactive activation loop conformations was also independent of phosphorylation. Importantly, we discovered that the presence of multiple enzyme conformations can lead to a plateau in the overall ligand K d, despite increasing affinity for the chosen target conformation, and modelled the conformational discrimination necessary for a conformation-promoting ligand.

SUBMITTER: Gilburt JAH 

PROVIDER: S-EPMC6461105 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

altmetric image

Publications

Ligand discrimination between active and inactive activation loop conformations of Aurora-A kinase is unmodified by phosphorylation.

Gilburt James A H JAH   Girvan Paul P   Blagg Julian J   Ying Liming L   Dodson Charlotte A CA  

Chemical science 20190304 14


Structure-based drug design is commonly used to guide the development of potent and specific enzyme inhibitors. Many enzymes - such as protein kinases - adopt multiple conformations, and conformational interconversion is expected to impact on the design of small molecule inhibitors. We measured the dynamic equilibrium between DFG-in-like active and DFG-out-like inactive conformations of the activation loop of unphosphorylated Aurora-A alone, in the presence of the activator TPX2, and in the pres  ...[more]

Similar Datasets

| S-EPMC3507260 | biostudies-literature
| S-EPMC3645566 | biostudies-literature
| S-EPMC7012767 | biostudies-literature
| S-EPMC5849412 | biostudies-literature
| S-EPMC5620369 | biostudies-literature
| S-EPMC5167317 | biostudies-literature
| S-EPMC2866269 | biostudies-literature
| S-EPMC3947352 | biostudies-literature
| S-EPMC7997955 | biostudies-literature
| S-EPMC3144914 | biostudies-literature